Differential expression of tandemly duplicated Enod8 genes in Medicago

Dickstein, Rebecca; Hu, Xuejun; Yang, Jong Oh; Ba, Lei; Coque, Laurent; Kim, Dong-Jin; Cook, Douglas R.; Yeung, Anthony T.

doi:10.1016/s0168-9452(02)00132-2

Cited by 17 publications

(34 citation statements)

References 33 publications

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“…Despite the apparent abundance of nodulationBecause the genetic markers used in this study are primarily expressed sequences or BAC clones that conassociated genes on linkage group 5, several nodulation genes (nodule expressed transcripts and phenotypically tain predicted genes, their position in the genome can be considered to provide a rough definition of the "gene mutant loci) are distributed elsewhere in the genome, including a cluster of ENOD8-like genes on linkage space" of M. truncatula. On the basis of cytogenetic analysis (Kulikova et al 2001), the structure of M. truncatula group 1 (Dickstein et al 2002), ENOD16 on linkage group 8, a cluster of apyrase genes on linkage group 7 chromosomes is apparently relatively simple, with condensed heterochromatic DNA in centromeric and peri- (Cohn et al 2001), and sunn, skl, dmi1, and dmi3 on linkage groups 4, 7, 2, and 8, respectively. centromeric islands, flanked by mostly euchromatic arms.…”

Section: Identification Of Bac Clones For Fish Analysismentioning

confidence: 99%

A Sequence-Based Genetic Map ofMedicago truncatulaand Comparison of Marker Colinearity withM. sativa

et al. 2004

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A core genetic map of the legume Medicago truncatula has been established by analyzing the segregation of 288 sequence-characterized genetic markers in an F 2 population composed of 93 individuals. These molecular markers correspond to 141 ESTs, 80 BAC end sequence tags, and 67 resistance gene analogs, covering 513 cM. In the case of EST-based markers we used an intron-targeted marker strategy with primers designed to anneal in conserved exon regions and to amplify across intron regions. Polymorphisms were significantly more frequent in intron vs. exon regions, thus providing an efficient mechanism to map transcribed genes. Genetic and cytogenetic analysis produced eight well-resolved linkage groups, which have been previously correlated with eight chromosomes by means of FISH with mapped BAC clones. We anticipated that mapping of conserved coding regions would have utility for comparative mapping among legumes; thus 60 of the EST-based primer pairs were designed to amplify orthologous sequences across a range of legume species. As an initial test of this strategy, we used primers designed against M. truncatula exon sequences to rapidly map genes in M. sativa. The resulting comparative map, which includes 68 bridging markers, indicates that the two Medicago genomes are highly similar and establishes the basis for a Medicago composite map.

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Section: Identification Of Bac Clones For Fish Analysismentioning

confidence: 99%

A Sequence-Based Genetic Map ofMedicago truncatulaand Comparison of Marker Colinearity withM. sativa

et al. 2004

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“…5A; Supplemental Table S1). Medtr1030230 is an MtENOD8 homolog found in the tandemly duplicated MtENOD8 gene cluster (Dickstein et al, 2002). None of the other similar SPs belong to proteins that are known to be expressed in symbiosomes or nodules; only one of the similar SPs belongs to a studied gene or protein: Medtr3g030400 encodes a gene corresponding to probeset Mtr.15106.1.…”

Section: Multiple Symbiosome-targeting Domains In Mtenod8mentioning

confidence: 99%

“…Stained membranes were cleared overnight in Tris-buffered saline (TBS) buffer containing 0.3% Tween 20 (TBS plus Tween 20). After being cleared of Coomassie Blue, membranes were blocked in blocking buffer (TBS plus Tween 20 containing 5% nonfat dry milk) at 25°C for 1 h. Membranes were immersed in blocking buffer containing 1:4,000 anti-ENOD8 antisera (Dickstein et al, 2002) for 1 h, rinsed for 5 min five times in TBS, incubated in blocking buffer containing 1:4,000 goat anti-rabbit IgG alkaline phosphatase conjugated antibody (Bethyl Laboratories), and rinsed for 5 min five times in TBS. Membranes were then equilibrated in 100 mM Tris pH 9.5 and developed with 5-bromo-4-chloro-3-indolyl phosphate/p-nitroblue tetrazolium chloride (Fisher Scientific).…”

Section: Plants Growth Conditions and Rhizobial Strainsmentioning

confidence: 99%

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Multiple Domains in MtENOD8 Protein Including the Signal Peptide Target It to The Symbiosome

et al. 2012

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Symbiotic nitrogen fixation occurs in nodules, specialized organs on the roots of legumes. Within nodules, host plant cells are infected with rhizobia that are encapsulated by a plant-derived membrane forming a novel organelle, the symbiosome. In Medicago truncatula, the symbiosome consists of the symbiosome membrane, a single rhizobium, and the soluble space between them, called the symbiosome space. The symbiosome space is enriched with plant-derived proteins, including the M. truncatula EARLY NODULIN8 (MtENOD8) protein. Here, we present evidence from green fluorescent protein (GFP) fusion experiments that the MtENOD8 protein contains at least three symbiosome targeting domains, including its N-terminal signal peptide (SP). When ectopically expressed in nonnodulated root tissue, the MtENOD8 SP delivers GFP to the vacuole. During the course of nodulation, there is a nodule-specific redirection of MtENOD8-SP-GFP from the vacuole to punctate intermediates and subsequently to symbiosomes, with redirection of MtENOD8-SP-GFP from the vacuole to punctate intermediates preceding intracellular rhizobial infection. Experiments with M. truncatula mutants having defects in rhizobial infection and symbiosome development demonstrated that the MtNIP/LATD gene is required for redirection of the MtENOD8-SP-GFP from the vacuoles to punctate intermediates in nodules. Our evidence shows that MtENOD8 has evolved redundant targeting sequences for symbiosome targeting and that intracellular localization of ectopically expressed MtENOD8-SP-GFP is useful as a marker for monitoring the extent of development in mutant nodules.

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“…A cluster of three ENOD8 is present in Medicago trunculata but one of them is probably a pseudogene for its lack of a 5' exon. The expression of the other two genes is located only in the nodule (Dickstein et al, 2002).…”

Section: Biology Of Nodules and Mycorrhizamentioning

confidence: 99%